Preliminary Study Of Kallistatin Gene On Male Infertility

Objective:Infertility has become an important disease in couples of childbearing age by affecting their health and lives.With the development of reproductive medicine and the conversion of people's traditional concepts,the problem of male infertility gained increasing attention.However,the reason of male infertility is very complex,the mechanism in pathological is not fully clear,no mention the special treatment at present.Which means exploring new therapeutic target is one of the hot spots.Spermatogenesis is one of the extremely important part of male reproductive function.The density,motility and morphology of Sperm determined its survivability and fertilizing capacity.The production of sperm is a series of complex and orderly process.Kallikrein-kinin system(KKS)is an important inflammatory regulation system in human body.Some studys found that it may play a regulatory role in the process of spermatogenesis.Kallistatin(KS),which is widely distributed in vivo and plays important roles in anti-oxidative stress,anti-fibrosis,diastolic blood vessels,anti-angiogenesis,anti-dampness and anti-angiogenesis in multiple systems and diseases.We found that kallistatin gene is related to rat spermatogenic function.kallistatin transgenic rat testicular tissue spermatogenic function was significantly enhanced.Therefore,we believe that kallistatin can also protect the male reproductive function through the local vasodilator effect,anti-oxidation,anti-apoptotic,anti-fibrosis and tissue damage repair.So our study will investigate the expression of kallistatin in testicular of different physiological age rats,and in seminal plasma and testicular of male infertility patients,to clarify the relationship with male infertility,and to investigate whether kallistatin could as a new therapeutic target for male infertility.Methods:Collecting 40 semen samples from male infertility patients and 10 normal semen samples,who treated in the Department of Reproductive Medicine of the First People's Hospital of Yunnan Province from October 2015 to February 2017,then detecting the expression of kallistatin protein in seminal plasma by ELISA,and investigate the correlation between kallistatin and male infertility;In the meantime,collecting 40 cases testicular tissue from different types of infertility patients(normal sperm,obstructive azoospermia,non-obstructive azoospermia and oligospermia),and 80 cases testicular tissue from rats at eight different physiological age(4 weeks,8 weeks,12 weeks,20 weeks,30 weeks,40 weeks,60 weeks,80 weeks),examing the expression of klliatatin by RT-PCR,Western blot and immunohistochemistry to further clarify the role of kallistatin in spermatogenesis.Results:The expression level of kallistatin protein in seminal plasma of male infertility patients was lower than that of normal control group,and the oligospermia's expression level of seminal plasma was lower than that of normal control group(42.5%).The expression levels of kallistatin in testicular testes of different types of male infertility were as follows:normal control group?obstructive azoospermia>severe oligospermia>non-obstructive azoospermia(P<0.05).Kallistatin was expressed in testicular testes of rats of different ages.The expression level of kallistatin was related to the age of the rats.The level of expression in the adolescent was lower than the control group.The expression level was increase with age,get a peak at 20weeks and keep it high till 40 weeks,and then decrease with age.The expression of kallistatin in testis tissue of patients with obstructive azoospermia was significantly lower than that of normal control group.The expression of Kallistatin protein in testis tissue was lower than that in normal control group,but the mRNA level was not different from normal control group.Conclusion:Kallistatin is associated with male infertility in this study.The expression level of kallistatin is reduced in the testes of male oligozoospermia patients and the patients with obstructive azoospermia,which suggesting that kallistatin may be associated with male spermatogenesis Function.The expression level of kallistatin in testes of different age rats is correlated with the age changing in rats,which is lower in young age and then increased gradually.The expression level of kallistatin reached the peak at the beginning of breeding period,and maintained to the maximum age of childbearing,finally decreased to a lower level with the increasing of age,which suggesting that kallistatin may play a regulatory role in the development and recession of male reproductive function.